Spectrophotometric approaches to develop a bacteriophage based assay to identify pathogens of poultry

鸡已被证明携带多种可以感染人类的细菌病原体。这些病原体中最重要和最流行的是产气荚膜梭菌、沙门氏菌和空肠弯曲杆菌。检测鸡携带的细菌种类并不简单，大多数测试依赖于培养、ELISA或分子方法，这些方法在特异性和选择性方面都存在问题。延迟识别病原体或错误识别病原体可能会导致鸡只接受无效的抗生素，这对细菌携带、动物福利和抗生素耐药性的发展都有影响。本项目的目的是开发一个基于噬菌体的原型诊断工具，以供兽医/农民使用，以快速有效地确定感染鸡的细菌病原体的身份。本项目将利用细菌与其特定噬菌体之间相互作用的特异性。我们的目标是识别每一种主要鸡病原体的噬菌体，并用一种独特的可检测标记标记它们或它们的尾部纤维蛋白。这将与一种专门开发的能够产生液滴的微流控设备相结合。然后，生物样本将被测试感兴趣的病原体，并将通过该设备。如果感染一种病原体的两个噬菌体记者附着在单个液滴中的特定细胞上，就会有该病原体特有的特定共定位信号。这一信号将被检测到，并使用光谱学来识别病原体。这种分析的发展将在许多情况下对细菌诊断产生重大影响，然而，直接的目标是解决当前和实际的影响食物链安全的问题，这项研究是导致翻译终点的强大科学的一个明显例子。

Chickens have been shown to carry a wide range of bacterial pathogens that can infect humans. The most important and prevalent of these pathogens are Clostridium perfringens, Salmonella spp, and Campylobacter jejuni,. Detecting what bacterial species the chickens carry is not straightforward and most tests rely on culture, ELISA or molecular approaches which all have problems with specificity and selectivity. Delay in identifying the pathogens, or misidentifying them can lead to the chickens receiving ineffective antibiotics, this has implications for bacterial carriage, animal welfare and the development of antibiotic resistance. The aim of this project is to develop a prototype phage based diagnostic tool that could be used by vets/farmers in order to establish the identity of bacterial pathogens that infect chickens in a quick and effective manner.This project will exploit the specificity of interaction between bacteria and their specific bacteriophages. We will aim to identify phages for each of the major chicken pathogens, and label them, or their tail fibre proteins, with a unique detectable marker. This will be in conjunction with a specifically developed microfluidic device able to produce droplets. Biological samples will then be tested for the pathogens of interest will be passed through the device. If two phage reporters that infect one pathogen attach to a particular cell within a single droplet, there will be a specific co-localised signal unique to that pathogen. This signal will be detected and the pathogen identified with the use of spectroscopy. The development of this assay would have a significant impact on bacterial diagnosis for many situations, however the immediate aim is to address a current and real problem affecting the safety of the food chain, this research is a clear example of strong science which leads to a translational endpoint.